Lymphocyte Antigen 6 (Ly6) Proteins: New Players in Chronic Pain

淋巴细胞抗原 6 (Ly6) 蛋白：慢性疼痛的新参与者

基本信息

批准号：
10784019
负责人：
Kimberly Gomez
金额：
$ 23.6万
依托单位：
NEW YORK UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-25 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10784019
关键词：
Action Potentials Affect Afferent Neurons Analgesics Behavior Behavioral Binding Biochemical Biological Assay Cell Communication Cell Maturation Cell Proliferation Cells Closure by clamp Data Dissociation Down-Regulation Electrophysiology (science)Evaluation Extracellular Protein Fingers Fluorescence-Activated Cell Sorting Gene Family Generations Genes Genetic Genus Lynx Glycoproteins Glycosylphosphatidylinositols Goals Human Immune In Situ Hybridization Injury Intrathecal Injections Invaded Knock-out Knockout Mice LY6E gene Lead Learning Ligation Link Lipids Lymphocyte antigen Macrophage Macrophage Activation Malignant Neoplasms Measures Membrane Modeling Molecular Mus Nerve Fibers Neurons Neuropathy Nicotinic Receptors Nociception Orthologous Gene Pain Pain Threshold Pathologic Patients Pattern Peptides Physiological Plasmids Play Population Production Protein Family Proteins Rattus Regulation Reporting Rodent Role Shaker potassium channel Signal Transduction Small Interfering RNA Snake Venoms Sodium Sodium Channel Specificity Spinal Cord Spinal Ganglia Stimulus Sumoylation Pathway Surface Syndrome Techniques Testing Time Toxin Transfection Up-Regulation Validation afferent nerve biophysical properties cancer type cell motility chronic neuropathic pain chronic pain collapsin response mediator protein-2 confocal imaging cytokine design extracellular gain of function mutation in vivo mechanical allodynia molecular sequence database nerve injury neuronal excitability neurotransmission neurotransmitter release novel overexpression pain behavior pain relief pain signal painful neuropathy patch clamp postsynaptic protein protein interaction response sensory system spared nerve trafficking transcriptome sequencing ubiquitin-protein ligase voltage

项目摘要

PROJECT SUMMARY Chronic pain is a pathological state where sensory neurons become hyperexcitable leading to nociceptive neurotransmission in the absence of a painful stimulus. Genetic and functional studies have established the voltage-gated sodium channel NaV1.7 as a major contributor to human pain signaling. Although the regulation of NaV1.7 is poorly understood, it is thought to involve mechanisms related to surface trafficking and regulation via protein-protein interactions. For instance, upregulation of SUMOylation of cytosolic collapsin response mediator protein 2 (CRMP2) and VGSC β-subunits, and downregulation of Nedd4-2 (a cytosolic E3 ubiquitin ligase) in a model of spared nerve injury induced chronic pain result in functional upregulation of NaV1.7 channels. My studies have identified lymphocyte antigen 6 (Ly6) proteins as a novel class of NaV1.7 channel modulators. Ly6 proteins are extracellular glycoproteins that are a hallmark of different types of cancer and have a role in cell proliferation, cell migration, cell–cell interactions, immune cell maturation, macrophage activation, and cytokine production. Ly6 proteins show structural resemblance to the three-fingered snake venom toxins that are known to modulate nicotinic acetylcholine receptors and voltage-gated sodium channels. Of these group of proteins, Ly6e and Lynx1 are common between humans and rodents. RNA-sequencing databases showed that both Ly6e and Lyn1 expression increases in different populations of dorsal root ganglia (DRG) neurons after nerve injury. Moreover, my preliminary findings show that: (i) there are higher Ly6e signal levels in DRGs after spared nerve injury (SNI); (ii) overexpression of Ly6e is associated with increased sodium currents in DRGs, and NaV1.7 currents in HEK cells; and (iii) intrathecal injection of Ly6e plasmid induces pain- like behaviors in naïve rats. These data led me to hypothesize that: (i) modulation of NaV1.7 channels by Ly6e and Lynx1 may lead to altered expression and activity of these channels during chronic pain, and that (ii) interfering with NaV1.7-Ly6e/Lynx1 interactions may relieve pain. Thus, the goals of this proposal are to investigate the role of Ly6 proteins (i) in sensory neurons and (ii) as molecular determinants of the altered functional activity of NaV1.7 channels in pain states. My Specific Aims are: (1) Investigate the physiological function(s) of Ly6e and Lynx1 in primary sensory neurons from rodents with and without nerve injury; (2) Identify and characterize Ly6e and Lynx1 as modulators of NaV1.7 channels in rodent and human dorsal root ganglia neurons; and (3) Identify specific interaction domain(s) in NaV1.7, Ly6e and Lynx1 and validation of in vivo target engagement. These studies are anticipated to advance our understanding of the role of Ly6e and Lynx1 in the sensory system, and their role as modulators of a key pain-associated voltage-gated sodium channel, NaV1.7.

项目摘要慢性疼痛是一种病理状态，感觉神经元变得过度兴奋，导致伤害感受在没有疼痛刺激的情况下，神经传递。遗传学和功能研究已经建立了电压门控钠通道NAV1.7是人为疼痛信号的主要因素。虽然调节 NAV1.7的理解很少，人们认为它涉及与表面运输和调节有关的机制蛋白质蛋白质相互作用。例如，胞质倒塌响应介质的sumoylation上调蛋白2（CRMP2）和VGSCβ-subunits，以及在A中的NEDD4-2（胞质E3泛素连接酶）的下调不幸的神经损伤引起的慢性疼痛导致NAV1.7通道的功能上调。我的研究已经确定淋巴细胞抗原6（Ly6）蛋白是新型NAV1.7通道调节剂。 LY6蛋白是细胞外糖蛋白，是不同类型癌症的标志，在细胞增殖，细胞迁移，细胞 - 细胞相互作用，免疫细胞成熟，巨噬细胞激活和细胞因子产生。 LY6蛋白与三指蛇毒毒素的结构相似已知可以调节烟碱乙酰胆碱受体和电压门控钠通道。的这些蛋白质Ly6e和lynx1在人类和啮齿动物之间很常见。 RNA测序数据库表明，在不同种群的背根神经节中LY6E和LYN1表达都会增加（DRG）神经损伤后神经元。此外，我的初步发现表明：（i）有更高的LY6E信号幸免的神经损伤后DRG的水平（SNI）；（ii）LY6E的过表达与钠增加有关 DRG中的电流和HEK细胞中的NAV1.7电流；（iii）鞘内注射ly6e质粒会诱导疼痛 - 就像天真大鼠的行为一样。这些数据导致我假设：（i）LY6E对NAV1.7通道的调制 lynx1可能导致这些通道在慢性疼痛期间的表达和活性改变，并且（ii）干扰NAV1.7-LY6E/LYNX1相互作用可能会挽救疼痛。那，该提议的目标是研究Ly6蛋白（I）在感觉神经元和（ii）中的作用作为改变的分子决定剂 NAV1.7通道在疼痛状态中的功能活性。我的具体目的是：（1）研究生理 Ly6e和lynx1的功能（s）在啮齿动物的原代感觉神经元中有或没有神经损伤的功能；（2）识别并将ly6e和lynx1描述为啮齿动物和人体背根神经节中NAV1.7通道的调节剂神经元；（3）在NAV1.7，LY6E和LYNX1中确定特定的交互域以及体内目标的验证订婚。预计这些研究将促进我们对Ly6e和lynx1在感觉系统及其作为关键疼痛相关电压门控钠通道的调节剂的作用，NAV1.7。